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Figure 2 | BMC Molecular Biology

Figure 2

From: The leukemia associated ETO nuclear repressor gene is regulated by the GATA-1 transcription factor in erythroid/megakaryocytic cells

Figure 2

Effects of 5'deletions on ETO promoter activity in erythroleukemia HEL and megakaryocytic MEG-01 cells. The following reporter constructs were examined: pGL3 -1820-259 (-1820), pGL3 -1326-259 (-1326), pGL3 -839-259 (-839), pGL3 -729-259 (-729), pGL3 -579-259 (-579), and pGL3 -429-259 (-429) after transfection into erythroid (HEL) and megakaryocytic (MEG-01) cell lines. Nucleotide +1 indicates the translational start site (ATG) and nucleotides 5'and 3' thereof have a "-" and "+" designation. The promoterless pGL3/basic and the pGL3/SV40-promoter are used as negative and positive control, respectively. Firefly and Renilla luciferase (internal standard) activities were assayed 24 h post-transfection. The luciferase activity is normalized against pGL3/SV40-promoter activity. The transcriptional activity of the full-length promoter was retained by the -729 to -259 bp region, which is therefore likely to contain the proximal ETO promoter. The -579 to -259/-429 to -171 bp regions showed lack of transcriptional activity. Firefly was normalized to Renilla luciferase as internal control for transfection efficiency and the results are given as adjusted Relative Luciferase Units (AdjRLU). Bars represent the mean of results from 3 to 5 separate transfections and the error bars show SEM.

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